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[目的]探讨一种新的获取海洋贝类基因组DNA的取样方法,为开展珍稀贝类的分子生物学研究提供参考。[方法]以文蛤、栉江瑶、翡翠贻贝、香港巨牡蛎、毛蚶为供试材料,以闭壳(合)肌全基因组DNA作为参考,采用常规酚-氯仿法抽提贝类贝腔液基因组DNA,使用生物光度计法、琼脂糖凝胶电泳、目的片段扩增测序验证其质量,并建立系统发育树检测其来源的真实可靠性。[结果]采用常规酚/氯仿基因组DNA提取方法获得的贝腔液DNA质量优于闭壳(合)肌DNA,蛋白质、多酚色素污染较少,完全满足目的片段扩增测序;系统发育进化树则证实了贝腔液并非外源污染。[结论]从贝腔液中获取基因组DNA是完全可行的,并可将其用于目的基因片段的扩增。
[Objective] The research aimed to explore a new sampling method for obtaining marine shellfish genomic DNA and provide a reference for the research of molecular biology of rare shellfish. [Method] The genomic DNAs of clams, combs, jade mussels, giant oysters and pomfret in Hong Kong were used as test materials, and the whole genomic DNA of clam shell was used as reference. The genomic DNA was verified by biophotometer method, agarose gel electrophoresis and sequencing of the target fragment, and its phylogenetic tree was established to test its authenticity. [Result] The results showed that the DNA quality of Beigu cavity liquid obtained by conventional phenol / chloroform genomic DNA extraction method was better than that of the closed shell muscle DNA, and the protein and polyphenol pigment were less contaminated, which fully met the target sequence. Phylogenetic tree Proved Beibei liquid is not exogenous pollution. [Conclusion] It is feasible to obtain genomic DNA from Beigu liquid and can be used to amplify the target gene fragment.